摘要
目的观察缺氧条件下7种不同类型细胞损伤程度,筛选对缺氧损伤敏感的细胞模型。方法选择CCD 841 CoN、GES-1、HUVEC、HEB、HEK-293、HBE和L02等7种不同组织来源的细胞,利用低氧工作站构建不同缺氧浓度(5%O_2和1%O_2)细胞缺氧模型,使用CCK-8和LDH方法检测细胞增殖及损伤情况。并用红景天(0.625μmol/L、2.5 mmol/L)和丁苯酞(1、2μmol/L)于缺氧开始时处理细胞,观察是否可改善缺氧细胞损伤。结果与21%O_2相比,1%O_2处理24 h时,HEB细胞增殖能力显著降低(P<0.05);与21%O_2相比,5%O_2与1%O_2处理24 h HEB、HUVEC、HBE、HEK-293、CCD 841 CoN、GES-1和L02细胞其上清中LDH酶活力均显著升高(P<0.05,P<0.01);在HEB细胞中,红景天(2.5 mmol/L)和丁苯酞(1μmol/L)处理可显著逆转低氧诱导的细胞增殖和细胞活力降低(P<0.05);红景天(2.5 mmol/L)和丁苯酞(1μmol/L)处理可显著逆转低氧诱导的LDH酶活力升高(P<0.05,P<0.01)。结论 HEB细胞对缺氧最为敏感,是较好的抗缺氧损伤药物效应研究细胞的模型。
Objective To observe hypoxic injuries in 7 cell lines derived from different human tissues to identify a cell model that is sensitive to acute hypoxic injury. Methods Seven cell lines derived from different human tissues, namely CCD 841 CoN, GES-1, HUVEC, HEB, HEK-293, HBE and L02, were exposed to different oxygen concentrations(1% and 5%) for 24 h using a hypoxia workstation. CCK-8 and LDH kits were used to assess the cell proliferation and cell injuries. The cells were then treated with rhodiola(0.625 μmol/L, 2.5 mmol/L) and 3-n-butylphthalide(1,2 μmol/L) at the start of hypoxic exposure to assess the protective effects of the drugs against hypoxic cell injuries. Results Compared with the cells cultured in normal oxygen concentration(21%), HEB cells exposed to 1% O_2 for 24 h showed significantly lowered proliferative activity(P<0.05). Hypoxic exposure to 5% or 1% O_2 for 24 h significantly increased LDH activity in the culture supernatant of HEB, HUVEC, HBE, HEK-293, CCD 841 CoN, GES-1 and L02 cells(P<0.05 or 0.01). In HEB cells cultured under hypoxic conditions, treatment with rhodiola and 3-n-butylphthalide significantly promoted the cell proliferation and improved the cell viability(P<0.05), and obviously reversed hypoxia-induced increase in LDH activity(P<0.05 or P<0.01). Conclusion Among the 7 cell lines tested in this study, HEB cells are the most sensitive to hypoxia and can serve as an ideal model for studying drug interventions of hypoxic cell injuries.
引文
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